A limited number of Y chromosome lineages is present in North American Holsteins.
Holsteins are the most numerous dairy cattle breed in North America and the breed has undergone intensive selection for improving milk production and conformation. Theoretically, this intensive selection could lead to a reduction of the effective population size and reduced genetic diversity. The objective of this study was to investigate the effective population size of the Holstein Y chromosome and the effects of limited Y chromosome lineages on male reproduction and the future of the breed. Paternal pedigree information of 62,897 Holstein bulls born between 1950 and 2013 in North America and 220,872 bulls evaluated by multiple-trait across-country genetic evaluations of Interbull (Uppsala, Sweden) were collected and analyzed. The results indicated that the number of Y chromosome lineages in Holsteins has undergone a dramatic decrease during the past 50 years because of artificial selection and the application of artificial insemination (AI) technology. All current Holstein AI bulls in North America are the descendants of only 2 ancestors (Hulleman and Neptune H) born in 1880. These 2 ancestral Y-lineages are continued through 3 dominant pedigrees from the 1960s; namely, Pawnee Farm Arlinda Chief, Round Oak Rag Apple Elevation, and Penstate Ivanhoe Star, with a contribution of 48.78, 51.06, and 0.16% to the Holstein bull population in the 2010s, respectively. The Y-lineage of Penstate Ivanhoe Star is almost eliminated from the breed. The genetic variations in the 2 ancestral Y-lineages were evaluated among 257 bulls by determining the copy number variations (CNV) of 3 Y-linked gene families: PRAMEY, HSFY, and ZNF280BY, which are spread along the majority (95%) of the bovine Y chromosome male-specific region (MSY). No significant difference was found between the 2 ancestral Y-lineages, although large CNV were observed within each lineage. This study suggests minimal genetic diversity on the Y chromosome in Holsteins and provides a starting point for investigating the effect of the extremely limited number of Y-lineages on male reproduction and other traits important for the future of the Holstein breed.


Comprehensive pathogen detection associated with four recurrent episodes of Kawasaki disease in a patient during a single year using next-generation sequencing.
Kawasaki disease (KD) is the most common multisystem vasculitis in childhood. Pathogens can be associated with the onset of KD. However, a lack of consistency prevails among reports about this disease. For this case of a 1-year-old boy with four recurrent episodes of KD within a year, we analysed profiles of pathogen reads in his serum and pharynx specimens using next-generation sequencing. Comparative analysis of the identified bacterial reads from serum samples found significant correlation of bacteria such as Streptococcus and Haemophilus spp. with the first and fourth episodes (R2 = 0.9506) before treatment. In the first convalescent phase, the number of Streptococcus spp. was reduced remarkably (P < 0.0001). From sequencing of the pharynx specimen from the fourth episode, a similar correlation was found with serum from the fourth episode (R2 = 0.6633). In this case, Streptococcus spp. may have been associated with onset of KD. Further studies must be undertaken to evaluate the putative association of micro-organism infection with KD pathogenesis.


A randomized controlled trial of household-based flocculant-disinfectant drinking water treatment for diarrhea prevention in rural Guatemala.
We conducted a study to determine if use of a new flocculant-disinfectant home water treatment reduced diarrhea. We randomly assigned 492 rural Guatemalan households to five different water treatment groups: flocculant-disinfectant, flocculant-disinfectant plus a customized vessel, bleach, bleach plus a vessel, and control. During one year of observation, residents of control households had 4.31 episodes of diarrhea per 100 person-weeks, whereas the incidence of diarrhea was 24% lower among residents of households receiving flocculant-disinfectant, 29% lower among those receiving flocculant-disinfectant plus vessel, 25% lower among those receiving bleach, and 12% lower among households receiving bleach plus vessel. In unannounced evaluations of home drinking water, free chlorine was detected in samples from 27% of flocculant-disinfectant households, 35% of flocculant-disinfectant plus vessel households, 35% of bleach households, and 43% of bleach plus vessel households. In a setting where diarrhea was a leading cause of death, intermittent use of home water treatment with flocculant-disinfectant decreased the incidence of diarrhea.


Vocal Characteristics of School-Aged Children With and Without Attention Deficit Hyperactivity Disorder.
The aim of this study was to describe the laryngeal anatomy, perceptual, acoustic, and aerodynamic vocal characteristics of school-aged children with and without Attention Deficit Hyperactivity Disorder (ADHD). The predisposition that children with ADHD have for laryngeal injuries are recurrent in nature and are more often than not overlooked as laryngitis. Previous studies have reported varied results on the prevalence rates of pediatric vocal fold nodules within the school-aged ADHD population. A static, two-group comparison was used in the study to investigate the clinical, perceptual, acoustic, and aerodynamic vocal characteristics of children between 7 and 9 years old with and without ADHD. The study replicated the protocol as executed by Barona-Lleo and Fernandez (2016) with additions. The Multidimensional Voice Program and the Voice Range Profile as additions to the assessment of vocal parameters were used with which comparable dysphonia severity index scores were calculated. Once-off clinical, perceptual, acoustic, and aerodynamic voice assessments were conducted on 20 age-gender matched participants (Control group mean age [months] = 98.80, standard deviation = 10.379; ADHD group mean age [months] = 108.00, standard deviation = 10.873). It was hypothesized that children with ADHD would have more hyperfunctional vocal characteristics; leading to laryngeal injuries, than their control group peers. Forty-five percent (n = 9) of the total sample population (both groups combined) had laryngeal pathology. Similar parent reported etiological voice symptoms and vocal habits were seen across both groups. Both groups performed similarly across both perceptual and aerodynamic voice assessments. Acoustically, the control group achieved significantly higher producible pitches than the ADHD group (P = 0.028) and were found to have more dysphonic dysphonia severity index scores than their ADHD group peers (P = 0.034). Prepubertal, school-aged children with or without ADHD may have similar vocal characteristics than previously thought. This is in support of the null hypothesis. The authors of the current study recommend that vocal screening in all school-aged children be carried out as an effective measure to monitor voice disorders in the pediatric population. Future research into larger sample sizes with this population with a special focus on the effect that central nervous system stimulants may have on the voice is recommended.


Localization and comparative nucleotide sequence analysis of the transforming domain in herpes simplex virus DNA containing repetitive genetic elements.
The 7.5-kilobase BamHI E fragment (BamHI-E) of herpes simplex virus type 2 (HSV-2) DNA (map position 0.533-0.583) encodes the 144-kDa subunit of ribonucleotide reductase and induces the neoplastic transformation of immortalized cell lines. To define the minimal transforming region of BamHI-E, a series of subclones were constructed that spanned the entire fragment. These subclones were assayed for focus formation in Rat-2 cells. Removal of the promoter region from the viral 144-kDa-protein gene left the transforming activity of DNA clones intact. A 481-bp Pst I-Sal I subclone of BamHI-E was capable of inducing focus formation and tumorigenic conversion. The nucleotide sequence of this fragment and the colinear nontransforming region of HSV-1 DNA was determined and compared. Striking differences were detected in the structure and organization of repeated sequence elements. Specifically, transforming HSV-2 DNA contains multiple regions of alternating purines and pyrimidines, G + C-rich sequences that are potential binding sites for transcription factor Sp1, and insertion-like sequence elements that are interrupted by base substitutions in nontransforming HSV-1 DNA. These results define a distinct transforming domain in HSV-2 DNA composed of repetitive elements implicated in gene rearrangement and activation.


[Influence of the minnow Phoxinus phoxinus L. sex on the composition of the parasite fauna and component community structure of parasites].
225 minnow specimens of the 2(+)-3+ age were collected and studied using the standard technique of the total parasitological dissection. The component parasite community in a period of studies was well-established. This state is characterized by maximum species richness and biomass values. The results showed that probable differences in the infection by parasites of minnow females and males could not be ignored and underestimated. Indeed, the differences in minnow of different sex's infestation by parasites may be absent or present. In some reservoirs females are stronger affected by the same kinds of parasites, in other reservoirs, males are affected stronger. In comparison to males, in females parasites are more commonly represented by single individuals. Discussed differences may manifest themselves only in a certain seasonal condition of a fish. The structure of parasitic communities in hosts of different sex is equal and almost identical to that obtained in mixed host samples.


Validation of the HIV Pre-exposure Prophylaxis Stigma Scale: Performance of Likert and Semantic Differential Scale Versions.
Stigma regarding HIV pre-exposure prophylaxis (PrEP) is commonly implicated as a factor limiting the scale-up of this highly effective HIV prevention modality. To quantify and characterize PrEP stigma, we developed and validated a brief HIV PrEP Stigma Scale (HPSS) among a group of 279 men who have sex with men (MSM). Scale development was informed by a theoretical model to enhance content validity. We assessed two scale versions, Semantic Differential and Likert, randomizing the order in which scales were presented to participants. Both scales demonstrated high internal consistency. The Likert scale had substantially better construct validity and was selected as the preferred option. Scale scores demonstrated construct validity through association with constructs of interest: healthcare distrust, HIV knowledge, perceived proportion of friends/partners on PrEP, perceived community evaluation of PrEP, and perceived effectiveness of PrEP. The scale accounted for 25% of the total variance in reported willingness to be on PrEP, indicating the substantial role PrEP stigma may have on decisions to initiate PrEP. Given increased efforts to roll-out PrEP, having a valid tool to determine the level and types of PrEP stigma in individuals, groups, and communities can help direct implementation plans, identify goals for stigma reduction, and monitor progress over time.


Drug shortages and the burden of access to care: a critical issue affecting patients with cancer.
Pharmaceutical drug shortages are multifaceted and complex problems that affect all aspects of health care, including patients, caregivers, healthcare providers, third-party payers, the pharmaceutical industry, and regulators. Drug shortages have increased significantly since 2000, which cause increases in healthcare costs and compromised patient care. New government regulations have led the U.S. Food and Drug Administration to focus efforts on updating policies and improving regulation of the pharmaceutical industry to limit and avoid drug shortages. This article discusses the current issues surrounding the pharmaceutical drug shortage and the implications for patients and healthcare providers. A review of the literature presents the multidimensional impact of the pharmaceutical drug shortage, and the analysis shows patients who are most burdened by drug shortages and have experienced substandard care, increased cost of care, and compromised quality of health care.


The occurrence and seasonal variation of aflatoxin B1 and zearalenone concentrations in poultry feeds.
The present study aimed to investigate the contamination of poultry feed with aflatoxin B1 and zearalenone at laying hen farms in Tehran suburbs. The poultry feed was selected from five laying hen farms. A total of 60 poultry feed samples were collected from each farm during four consecutive seasons, from spring to winter of 2021. High-performance liquid chromatography was used to determine the amount of aflatoxin B1 and zearalenone. The mean aflatoxin B1 and zearalenone concentrations in various seasons showed significant differences (p<0.01). The highest reported aflatoxin concentration was in winter, with a mean concentration of 1366.53±77.85 ng/kg. The lowest concentrations were reported in autumn and summer, indicating a significant difference (p<0.01). The highest concentration of zearalenone was reported in summer, with a mean concentration of 150.72±10.35 μg/kg. The lowest concentration was reported in winter, with a mean concentration of 22.87±10.35 μg/kg, indicating a statistically significant difference (p<0.01). Overall, the concentrations of aflatoxin B1 and zearalenone toxins significantly differed in various poultry farms. The poultry farm D had the highest aflatoxin contamination with a mean concentration of 648.08±59.89 ng/kg. Poultry farms A, B, and C had the highest zearalenone concentrations with mean concentrations of 125.17±20.61, 96.04±20.61, and 99.49±20.61 μg/kg, respectively. Autumn was the only season showing significant differences regarding zearalenone toxin concentration in poultry farms.


Pulmonary arterial pressure at rest and during exercise in chronic mountain sickness: a meta-analysis.
Up to 10% of the more than 140 million high-altitude dwellers worldwide suffer from chronic mountain sickness (CMS). Patients suffering from this debilitating problem often display increased pulmonary arterial pressure (PAP), which may contribute to exercise intolerance and right heart failure. However, there is little information on the usual PAP in these patients.We systematically reviewed and meta-analysed all data published in English or Spanish until June 2018 on echocardiographic estimations of PAP at rest and during mild exercise in CMS patients.Nine studies comprising 287 participants fulfilled the inclusion criteria. At rest, the point estimate from meta-analysis of the mean systolic PAP was 27.9 mmHg (95% CI 26.3-29.6 mmHg). These values are 11% (+2.7 mmHg) higher than those previously meta-analysed in apparently healthy high-altitude dwellers. During mild exercise (50 W) the difference in mean systolic PAP between patients and high-altitude dwellers was markedly more accentuated (48.3 versus 36.3 mmHg) than at rest.These findings indicate that in patients with CMS PAP is moderately increased at rest, but markedly increased during mild exercise, which will be common with activities of daily living.


Single-step ambient-air synthesis of graphene from renewable precursors as electrochemical genosensor.
Thermal chemical vapour deposition techniques for graphene fabrication, while promising, are thus far limited by resource-consuming and energy-intensive principles. In particular, purified gases and extensive vacuum processing are necessary for creating a highly controlled environment, isolated from ambient air, to enable the growth of graphene films. Here we exploit the ambient-air environment to enable the growth of graphene films, without the need for compressed gases. A renewable natural precursor, soybean oil, is transformed into continuous graphene films, composed of single-to-few layers, in a single step. The enabling parameters for controlled synthesis and tailored properties of the graphene film are discussed, and a mechanism for the ambient-air growth is proposed. Furthermore, the functionality of the graphene is demonstrated through direct utilization as an electrode to realize an effective electrochemical genosensor. Our method is applicable to other types of renewable precursors and may open a new avenue for low-cost synthesis of graphene films.


Levitated droplet dye laser.
We present the first observation, to our knowledge, of lasing from a levitated, dye droplet. The levitated droplets are created by computer controlled pico-liter dispensing into one of the nodes of a standing ultrasonic wave (100 kHz), where the droplet is trapped. The free hanging droplet forms a high quality optical resonator. Our 750 nL lasing droplets consist of Rhodamine 6G dissolved in ethylene glycol, at a concentration of 0.02 M. The droplets are optically pumped at 532 nm light from a pulsed, frequency doubled Nd:YAG laser, and the dye laser emission is analyzed by a fixed grating spectrometer. With this setup we have achieved reproducible lasing spectra in the visible wavelength range from 610 nm to 650 nm. The levitated droplet technique has previously successfully been applied for a variety of bio-analytical applications at single cell level. In combination with the lasing droplets, the capability of this high precision setup has potential applications within highly sensitive intra-cavity absorbance detection.


A randomized clinical trial to compare the efficacy of three treatment regimens along with footcare in the morbidity management of filarial lymphoedema.
The progression of lymphoedema to elephantiasis associated with increased incidence of episodic adeno-lymphangitis (ADL) is of great concern, as it causes physical suffering, permanent disability and economic loss to lymphatic filariasis patients. This randomized clinical trial aimed to assess the efficacy in terms of reduction of oedema and ADL frequency of three treatment regimens among lymphoedema patients from Orissa, India. The regimens were: (I) oral penicillin--one tablet of 800 000 U penicillin G potassium twice daily for 12 days--repeated every 3 months for 1 year; (II) diethylcarbamazine--6 mg/kg bodyweight for 12 days-repeated every 3 months for 1 year; and (III) topical antiseptic, i.e. betadine ointment. Foot care was part of all regimens. All three drug regimens are efficacious in reducing oedema and frequency of ADL episodes. Although the efficacy was slightly higher in regimen I, the difference was not significant. About half of all patients had reduced oedema after the 90 days of treatment, with oedema reduction of 75-100% in 20%. A major proportion of the remaining patients had oedema reduced by less than 25%. The proportion of people whose oedema reduced was slightly but not significantly lower in regimen II. anova revealed that lymphoedema reduction varied according to grade; being greatest at grade 1 lymphoedema, followed by grade 2. All three regimens significantly reduced ADL frequency after 1 year of treatment. This may be because of foot care as well as use of antibiotics. The estimated costs of treatment per patient for a period of 3 months are US$2.4, 1.5 and 4.0 respectively for regimen I, II and III. Thus affordable treatments with simple antibiotics and foot care can give substantial relief to the patients and reverse early lymphoedema.


Ultrastructure of human tracheal smooth muscle from subjects with asthma and nonasthmatic subjects. Standardized methods for comparison.
A characteristic feature of asthma is exaggerated airway narrowing, termed airway hyper-responsiveness (AHR) due to contraction of airway smooth muscle (ASM). Although smooth muscle (SM)-specific asthma susceptibility genes have been identified, it is not known whether asthmatic ASM is phenotypically different from nonasthmatic ASM in terms of subcellular structure or mechanical function. The present study is the first to systematically quantify, using electron microscopy, the ultrastructure of tracheal SM from subjects with asthma and nonasthmatic subjects. Methodological details concerning tissue sample preparation, ultrastructural quantification, and normalization of isometric force by appropriate morphometric parameters are described. We reasoned that genetic and/or acquired differences in the ultrastructure of asthmatic ASM could be associated with functional changes. We recently reported that asthmatic ASM is better able to maintain and recover active force generation after length oscillations simulating deep inspirations. The present study was designed to seek structural evidence to account for this observation. Contrary to our hypotheses, no significant qualitative or quantitative differences were found in the subcellular structure of asthmatic versus nonasthmatic tracheal SM. Specifically, there were no differences in average SM cell cross-sectional area; fraction of the cell area occupied by nonfilamentous area; amounts of mitochondria, dense bodies, and dense plaques; myosin and actin filament densities; basal lamina thickness; and the number of microtubules. These results indicate that functional differences in ASM do not necessarily translate into observable structural changes.


Hypoglycemia but not hyperglycemia induces rapid changes in pancreatic beta-cell gene transcription.
The purpose of these studies was to quantify several mRNAs expressed specifically in pancreatic islet cells and known or postulated to be important for insulin release after acute well defined alterations in levels of plasma glucose. Glucose levels were maintained at 50, 120, or 180 mg/dl (2.8, 6.7, or 10 mM) for 3 h in conscious unrestrained rats. Hypoglycemia (for 3 h) caused significant decreases in pancreatic content of mRNAs for insulin 2 and GLUT-2 to 55 and 34% of control values, respectively. There were no significant changes in insulin 1, amylin, glucokinase, or glucagon mRNAs. Unprocessed insulin 1 and 2 mRNA precursors were decreased to 17 and 10% of levels in controls, consistent with effects of short-term hypoglycemia on new mRNA synthesis. Hyperglycemia (for 3 h) caused no increase in pancreatic content of any mRNA measured. To discriminate between effects of hypoglycemia and hyperinsulinemia in the hypoglycemic animals, rats were made hypoglycemic by infusion with etomoxir, a carnitine palmitoyltransferase I inhibitor that lowers glucose in the fasted (glycogen-depleted) state by inhibiting hepatic gluconeogenesis. A single dose of this agent caused a decrease in glucose from 120 mg/dl (6.7 mM) to 80 mg/dl (4.4 mM) and significantly decreased insulin mRNA and pre-mRNA. These results are consistent with the hypothesis that glucose modulates islet cell gene transcription directly. They indicate that the range of glucose concentrations that modulate gene transcription differs from the levels of glucose that alter both insulin biosynthetic and secretion rates.


Relationships between bullying and violence among US youth.
To determine the extent to which bullying and being bullied, both in and away from school, is associated with involvement in violent behavior. A US representative cross-sectional sample of 15 686 students in grades 6 through 10 in public and private schools completed the World Health Organization's Health Behaviour in School-aged Children survey in 1998. Self-report of weapon carrying, weapon carrying in school, physical fighting, and being injured in a physical fight. Involvement in each of the violence-related behaviors ranged from 13% to 23% of boys and 4% to 11% of girls. Bullying others and being bullied were consistently related to each violence-related behavior for both boys and girls. Greater odds of involvement occurred with bullying others than being bullied, and greater odds of involvement occurred with bullying that took place away from school than that occurring in school. For example, the adjusted odds ratio for weapon carrying associated with being bullied in school weekly was 1.5, for bullying others in school 2.6, for being bullied away from school 4.1, and for bullying others away from school 5.9. Bullying should not be considered a normative aspect of youth development, but rather a marker for more serious violent behaviors, including weapon carrying, frequent fighting, and fighting-related injury.


Obesity, Even in the Metabolically Healthy, Increases the Risk of Poor Physical Performance: A Cross-Sectional Study of Older People in a Chinese Community.
We examined the association between obesity and physical performance under different metabolic status. The sample included 1395 Chinese community-dwelling participants (mean age, 71.88 ± 5.87 years; 40.9% men). Being metabolically healthy was defined as having the presence of < 3 of 5 components of metabolic syndrome (MetS); obesity was defined as having a BMI > 28 kg/m2. Participants were divided into four groups based on BMI (non-obese/obese) and metabolic health (healthy/unhealthy). Physical performance was measured by grip strength, 4-m walking speed, and the timed up and go test (TUGT). After multiple adjustments, compared with metabolically healthy non-obese group, the metabolically unhealthy obese group showed lower relative grip strength, lower 4-m walking speed, and higher TUGT (P all < 0.05), and only relative grip strength of the metabolically healthy obese group was significantly lower than that of metabolically healthy non-obese (P < 0.01). Relative grip strength was negatively associated with impaired fasting glucose (β = -0.071), elevated triglycerides (β = -0.062), abdominal obesity (β = -0.230) and general obesity (β = -0.225) (P all < 0.01). Walking speed and TUGT were only associated with general obesity, rather than other metabolic components. The associations of MetS with physical performance were mainly driven by abdominal obesity. Even in those who are metabolically healthy, obesity (especially general obesity) increases the risk of poor physical performance. Elderly people with general obesity and MetS, whether in combination or alone, have an increased risk of muscle dysfunction, and that combination produces a higher risk of impaired mobility.


Ultrasound, infrared and its assisted technology, a promising tool in physical food processing: A review of recent developments.
Traditional food processing techniques can no longer meet the ever increasing demand for high quality food across the globe due to its low process efficiency, high energy consumption and low product yield. This review article is focused on the mechanism and application of Infrared (IR) and ultrasound (US) technologies in physical processing of food. We herein present the individual use of IR and US (both mono-frequency and multi-frequency levels) as well as IR and US supported with other thermal and non-thermal technologies to improve their food processing performance. IR and US are recent thermal and non-thermal technologies which have now been successfully used in food industries to solve the demerits of conventional processing technologies. These environmentally-friendly technologies are characterized by low energy consumption, reduced processing time, high mass-transfer rates, better nutrient retention, better product quality, less mechanical damage and improved shelf life. This work could be, with no doubt, useful to the scientific world and food industries by providing insights on recent advances in the use of US and IR technology, which can be applied to improve food processing technologies for better quality and safer products.


Is Enhancement the Price of Prevention in Human Gene Editing?
New gene-editing tools challenge conventional policy proscriptions of research aimed at either human germline gene editing or human enhancement by potentially lowering technical barriers to both kinds of intervention. Some recent gene-editing reports have begun to take up the prospect of germline editing, but most experts are in broad agreement that research should prioritize medical applications over attempts to enhance human traits. However, there is little consensus about what counts as human enhancement in this context, or how to deal with the issues it flags. Moreover, several influential reports interpret medical applications to include disease prevention as well as treatment as a goal for gene-editing research. This challenges the current policy consensus because using gene editing to prevent disease would incidentally facilitate human enhancement applications in a variety of ways. If such research efforts are penalized by policy concerns about enhancement, then their preventive health benefits could be lost. To avoid being caught off guard by such challenges, science policy makers will need to think more carefully about what "prevention" might mean in the gene-editing context, and develop research governance that can anticipate and address the human enhancement concerns it will raise. To accomplish the latter, the scope of policy making will need to expand from its narrow focus on human clinical trials to engage with basic researchers driving the translational pipeline toward preventive gene editing and the science policy makers who have to address its "off-label" uses.


Impact of hypoxia on chemoresistance of mesothelioma mediated by the proton-coupled folate transporter, and preclinical activity of new anti-LDH-A compounds.
Expression of proton-coupled folate transporter (PCFT) is associated with survival of mesothelioma patients treated with pemetrexed, and is reduced by hypoxia, prompting studies to elucidate their correlation. Modulation of glycolytic gene expression was evaluated by PCR arrays in tumour cells and primary cultures growing under hypoxia, in spheroids and after PCFT silencing. Inhibitors of lactate dehydrogenase (LDH-A) were tested in vitro and in vivo. LDH-A expression was determined in tissue microarrays of radically resected malignant pleural mesothelioma (MPM, N = 33) and diffuse peritoneal mesothelioma (DMPM, N = 56) patients. Overexpression of hypoxia marker CAIX was associated with low PCFT expression and decreased MPM cell growth inhibition by pemetrexed. Through integration of PCR arrays in hypoxic cells and spheroids and following PCFT silencing, we identified the upregulation of LDH-A, which correlated with shorter survival of MPM and DMPM patients. Novel LDH-A inhibitors enhanced spheroid disintegration and displayed synergistic effects with pemetrexed in MPM and gemcitabine in DMPM cells. Studies with bioluminescent hypoxic orthotopic and subcutaneous DMPM athymic-mice models revealed the marked antitumour activity of the LDH-A inhibitor NHI-Glc-2, alone or combined with gemcitabine. This study provides novel insights into hypoxia/PCFT-dependent chemoresistance, unravelling the potential prognostic value of LDH-A, and demonstrating the preclinical activity of LDH-A inhibitors.


Cardiovascular risks in prediabetes: preliminary data on "vasculopathy triad".
Subclinical cardiovascular disease is inherent in complications of diabetes mellitus. It occurs before the obvert manifestation of cardiovascular disease complication in diabetes, and involves vasculopathy triad or three major vascular events comprising stasis, endothelial dysfunction, and atherothrombosis. This study was to examine evidence of vasculopathy triad in prediabetes, biomarkers of stasis, endothelial dysfunction, and atherothrombosis in prediabetes were compared with apparently healthy group. Eighty-one participants with results for plasma D-dimer, homocysteine, and whole blood viscosity were selected from a research database. The participants consisted of control (n = 44) and prediabetes (n = 37) based on clinical history and laboratory results. Multivariate analysis shows a significantly higher level of vasculopathy in prediabetes than in the control group (P > 0.0001). Blood viscosity (P < 0.04) and homocysteine (P < 0.03) are significantly higher in prediabetes than in controls. Average levels for plasma D-dimer are also higher in prediabetes than in control, but not statistically significant in this particular analysis. This study suggests a novel application of known idea, vasculopathy triad that could be used for assessment of subclinical cardiovascular disease in prediabetes.


Cav1.4alpha1 subunits can form slowly inactivating dihydropyridine-sensitive L-type Ca2+ channels lacking Ca2+-dependent inactivation.
The neuronal L-type calcium channels (LTCCs) Cav1.2alpha1 and Cav1.3alpha1 are functionally distinct. Cav1.3alpha1 activates at lower voltages and inactivates more slowly than Cav1.2alpha1, making it suitable to support sustained L-type Ca2+ inward currents (ICa,L) and serve in pacemaker functions. We compared the biophysical and pharmacological properties of human retinal Cav1.4alpha1 using the whole-cell patch-clamp technique after heterologous expression in tsA-201 cells with other L-type alpha1 subunits. Cav1.4alpha1-mediated inward Ba2+ currents (IBa) required the coexpression of alpha2delta1 and beta3 or beta2a subunits and were detected in a lower proportion of transfected cells than Cav1.3alpha1. IBa activated at more negative voltages (5% activation threshold; -39mV; 15 mm Ba2+) than Cav1.2alpha1 and slightly more positive than Cav1.3alpha1. Voltage-dependent inactivation of IBa was slower than for Cav1.2alpha1 and Cav1.3alpha1( approximately 50% inactivation after 5 sec; alpha2delta1 + beta3 coexpression). Inactivation was not increased with Ca2+ as the charge carrier, indicating the absence of Ca2+-dependent inactivation. Cav1.4alpha1 exhibited voltage-dependent, G-protein-independent facilitation by strong depolarizing pulses. The dihydropyridine (DHP)-antagonist isradipine blocked Cav1.4alpha1 with approximately 15-fold lower sensitivity than Cav1.2alpha1 and in a voltage-dependent manner. Strong stimulation by the DHP BayK 8644 was found despite the substitution of an otherwise L-type channel-specific tyrosine residue in position 1414 (repeat IVS6) by a phenylalanine. Cav1.4alpha1 + alpha2delta1 + beta channel complexes can form LTCCs with intermediate DHP antagonist sensitivity lacking Ca2+-dependent inactivation. Their biophysical properties should enable them to contribute to sustained ICa,L at negative potentials, such as required for tonic neurotransmitter release in sensory cells and plateau potentials in spiking neurons.


Guest-Host Relationship of Cyclodextrin and Its Pharmacological Benefits.
Many methods, including solid dispersion, micellization, and inclusion complexes, have been employed to increase the solubility of potent drugs. Beta-cyclodextrin (βCD) is a cyclic oligosaccharide consisting of seven glucopyranoside molecules, and is a widely used polymer for formulating soluble inclusion complexes of hydrophobic drugs. The enzymatic activity of Glycosyltransferase or α-amylase converts starch or its derivatives into a mixture of cyclodextrins. The βCD units are characterized by α-(1-4) glucopyranose bonds. Cyclodextrins possess certain properties that make them very distinctive because of their toroidal or truncated cage-like supramolecular configurations with multiple hydroxyl groups at each end. This allowed them to encapsulate hydrophobic compounds by forming inclusion complexes without losing their solubility in water. Chemical modifications and newer derivatives, such as methylated βCD, more soluble hydroxyl propyl methyl βCD, and sodium salts of sulfobutylether-βCD, known as dexolve® or captisol®, have envisaged the use of CDs in various pharmaceutical, medical, and cosmetic industries. The successful inclusion of drug complexes has demonstrated improved solubility, bioavailability, drug resistance reduction, targeting, and penetration across skin and brain tissues. This review encompasses the current applications of β-CDs in improving the disease outcomes of antimicrobials and antifungals as well as anticancer and anti-tubercular drugs.


Impact of sex, androgens, and prostate size on C57BL/6J mouse urinary physiology: functional assessment.
Laboratory mice are used to identify causes of urinary dysfunction including prostate-related mechanisms of lower urinary tract symptoms. Effective use of mice for this purpose requires a clear understanding of molecular, cellular, anatomic, and endocrine contributions to voiding function. Whether the prostate influences baseline voiding function has not been specifically evaluated, in part because most methods that alter prostate mass also change circulating testosterone concentrations. We performed void spot assay and cystometry to establish a multiparameter "baseline" of voiding function in intact male and female 9-wk-old (adult) C57BL/6J mice. We then compared voiding function in intact male mice to that of castrated male mice, male (and female) mice treated with the steroid 5α-reductase inhibitor finasteride, or male mice harboring alleles (Pbsn4cre/+; R26RDta/+) that significantly reduce prostate lobe mass by depleting prostatic luminal epithelial cells. We evaluated aging-related changes in male urinary voiding. We also treated intact male, castrate male, and female mice with exogenous testosterone to determine the influence of androgen on voiding function. The three methods used to reduce prostate mass (castration, finasteride, and Pbsn4cre/+; R26RDta/+) changed voiding function from baseline but in a nonuniform manner. Castration feminized some aspects of male urinary physiology (making them more like intact female mice) while exogenous testosterone masculinized some aspects of female urinary physiology (making them more like intact male mice). Our results provide evidence that circulating testosterone is responsible in part for baseline sex differences in C57BL/6J mouse voiding function while prostate lobe mass in young, healthy adult mice has a lesser influence.


Graduate nursing concentration in women's health at the University of Illinois at Chicago.
The Graduate Nursing Concentration in Women's Health was formalized in the 1980s as a program of study that emphasizes nursing theory, research, and practice related to women's health. We describe the purposes of the program and the assumptions underlying the care of women as well as the curriculum and characteristics desired of graduates. In this program, the goal of advancing women's health is balanced by the desire to create a productive, educational environment that facilitates the personal construction of knowledge by learners.


Prenatal depression and birth mode sequentially mediate maternal education's influence on infant sleep duration.
Sleep duration is critical to growth, learning, and immune function development in infancy. Strategies to ensure that national recommendations for sleep duration in infants are met require knowledge of perinatal factors that affect infant sleep. To investigate the mechanistic pathways linking maternal education and infant sleep. An observational study was conducted on 619 infants whose mothers were enrolled at the Edmonton site of the CHILD birth cohort. Infant sleep duration at three months was assessed using the Brief Infant Sleep Questionnaire. Maternal education was collected via maternal report. Prenatal and postnatal depression scores were obtained from the 20-item Center for Epidemiologic Studies Depression Scale (CES-D). Birth records and maternal report were the source of covariate measures. Mediation analysis (PROCESS v3.0) was used to examine the indirect effects of maternal education on infant sleep duration mediated through prenatal depression and birth mode. At three months of age, infants slept on average 14.1 h. Lower maternal education and prenatal depression were associated with significantly shorter infant sleep duration. Emergency cesarean section birth was associated with 1-hour shorter sleep duration at three months compared to vaginal birth [without intrapartum antibiotic prophylaxis] (β: -0.99 h; 95% CI: -1.51, -0.48). Thirty percent of the effect of lower maternal education on infant total sleep duration was mediated sequentially through prenatal depression and birth mode (Total Indirect Effects: -0.12, 95% CI: -0.22, -0.03, p < 0.05). Prenatal depression and birth mode sequentially mediate the effect of maternal education on infant sleep duration.


Physical and physiological attributes of female volleyball players--a review.
The main objective of this article was to review a series of studies (n = 31) on physical attributes, physiological attributes, and on-court performances of female volleyball players. Empirical and practical knowledge emerging from studies on training-related issues in volleyball, such as body mass, fat-free mass, aerobic profile, strength, and agility and speed, should be integrated and applied when planning annual training programs for volleyball players. Based on our review, it was found that (a) players of a higher skill level are taller, somewhat heavier, and have higher vertical jump values than players of a lower level; (b) the aerobic profile of female volleyball players is similar to that of female basketball players; (c) ballistic resistance training can increase vertical jump values in female volleyball players; and (d) preseason conditioning should be conducted to prevent fatigue and reduced performance at the beginning of the season. Among the research concerns discussed in the article are that there is a lack data for on-court performance and time-motion analysis in female volleyball players and that more experimental/manipulative studies are needed to examine the effectiveness of different training programs on physiological attributes of female volleyball players. Two practical implications are suggested for volleyball and strength and conditioning coaches: (a) functional and nonfunctional overreaching should be carefully monitored when planning strength and conditioning programs, and (b) volleyball programs should include ballistic-type training.


Towards a scientific concept of free will as a biological trait: spontaneous actions and decision-making in invertebrates.
Until the advent of modern neuroscience, free will used to be a theological and a metaphysical concept, debated with little reference to brain function. Today, with ever increasing understanding of neurons, circuits and cognition, this concept has become outdated and any metaphysical account of free will is rightfully rejected. The consequence is not, however, that we become mindless automata responding predictably to external stimuli. On the contrary, accumulating evidence also from brains much smaller than ours points towards a general organization of brain function that incorporates flexible decision-making on the basis of complex computations negotiating internal and external processing. The adaptive value of such an organization consists of being unpredictable for competitors, prey or predators, as well as being able to explore the hidden resource deterministic automats would never find. At the same time, this organization allows all animals to respond efficiently with tried-and-tested behaviours to predictable and reliable stimuli. As has been the case so many times in the history of neuroscience, invertebrate model systems are spearheading these research efforts. This comparatively recent evidence indicates that one common ability of most if not all brains is to choose among different behavioural options even in the absence of differences in the environment and perform genuinely novel acts. Therefore, it seems a reasonable effort for any neurobiologist to join and support a rather illustrious list of scholars who are trying to wrestle the term 'free will' from its metaphysical ancestry. The goal is to arrive at a scientific concept of free will, starting from these recently discovered processes with a strong emphasis on the neurobiological mechanisms underlying them.


Nanoparticle based fluorescence resonance energy transfer (FRET) for biosensing applications.
In the past few decades, Förster resonance energy transfer (FRET) has been used as a powerful tool for providing nanoscale information in many biosensing and bioanalysis applications. The performance of FRET assays is mainly dependent on the design of donor and acceptor pairs. Recently, a series of nanoparticles start to be used in FRET assays including semiconductor quantum dots (QDs), graphene quantum dots (GQDs), upconversion nanoparticles (UCNPs), gold nanoparticles (AuNPs) and graphene oxide (GO). The rapid pace of development in nanoparticles provides a lot of opportunities to revolutionize FRET techniques. Many nanoparticle based FRET assays have also been developed for various biosensing applications with higher sensitivity and better stability compared with traditional organic fluorophore based FRET assays. This article reviews the recent progress of nanoparticle FRET assays and their applications in biosensing area.


Anion detection by fluorescent Zn(II) complexes of functionalized polyamine ligands.
The Zn(2+) coordination chemistry and luminescent behavior of two ligands constituted by an open 1,4,7-triazaheptane chain functionalized at both ends with 2-picolyl units and either a methylnaphthyl (L1) or a dansyl (L2) fluorescent unit attached to the central amino nitrogen are reported. The fluorescent properties of the ZnL1(2+) and ZnL2(2+) complexes are then exploited toward detection of anions. L1 in the pH range of study has four protonation constants. The fluorescence emission from the naphthalene fluorophore is quenched either at low or at high pH values leading to an emissive pH window centered around pH = 5. In contrast, in L2 the fluorescence emission from the dansyl unit occurs only at basic pH values. In the case of L1, a red-shifted band appearing in the visible region was assigned to an exciplex emission involving the naphthalene and the tertiary amine of the polyamine chain. L1 forms Zn(2+) mononuclear complexes of ZnH(p)L1((p+2)+) stoichiometry with p = 1, 0, -1. Formation of the ZnL1(2+)species produces a strong enhancement of the L1 luminescence leading to an extended emissive pH window from pH = 5 to pH = 9. Addition of several anions to this last complex leads to a partial quenching effect. On the contrary, the fluorescence emission of L2 is partially quenched upon complexation with Zn(2+) in the same pH window (5 < pH < 9). The lower stability of ZnL2(2+) with respect to ZnL1(2+) suggests a lack of involvement of the sulfonamide group in the first coordination sphere. However, there is spectral evidence for an interesting photoinduced binding of the sulfonamide nitrogen to Zn(2+). While addition of diphosphate, triphosphate, citrate, and D,L-isocitrate to a solution of ZnL2(2+) restores the fluorescence emission of the system (lambda max ca. 600 nm), addition of phosphate, chloride, iodide, and cyanurate do not produce any significant change in fluorescence. Moreover, this system would permit one to differentiate diphosphate and triphosphate over citrate and d, l-isocitrate because the fluorescence enhancement observed upon addition of the first anions is much sharper. The ZnL2(2+) complex and its mixed complexes with diphosphate, triphosphate, citrate, and D,L-isocitrate have been characterized by (1)H, (31)P NMR, and Electrospray Mass Spectrometry.


Vasoproliferation and antiproliferative treatment options in pulmonary arterial hypertension.
New treatment options have improved the prognosis of patients with pulmonary arterial hypertension. However, drugs such as prostanoids, PDE5 inhibitors and endothelin receptor antagonists mainly act as vasodilating agents. Recently, it has become clear that pulmonary arterial hypertension is an inflammatory and vasoproliferative disease. Therefore new anti-inflammatory and antiproliferative treatments are needed. This review will focus on the pathogenesis of inflammation and vasoproliferation in pulmonary hypertension. In addition, an overview on possible new antiinflammatory and antiproliferative drugs in pulmonary hypertension (e.g. Rho-kinase inhibitors, imatinib mesylate. HMG-CoA reductase inhibitors) will be given along with recent patents.


Cholinergic projections from the basal forebrain of rat to the amygdala.
Cholinergic afferents to the amygdala from the basal forebrain were studied using di-isopropyl fluorophosphate-AChE histochemistry in combination with retrograde tracing using various fluorescent dyes. Cells sending their axons to the amygdala and staining intensely for AChE were located mainly in the nucleus of the substantia innominata. They also were found in the ventral part of the globus pallidus, the horizontal limb of the nucleus tractus diagonalis Broca, and the nucleus interstitialis ansae lenticularis. A correspondence was established between these cells and cells staining for choline acetyltransferase by immunohistochemistry in both distribution and morphology. Non-cholinergic neurons which send their axons to the amygdala also were found in the substantia innominata complex.


Performance, ruminal changes, behaviour and welfare of growing heifers fed a concentrate diet with or without barley straw.
Two experiments were conducted to ascertain the effects of feeding an all-concentrate diet to growing heifers on performance, ruminal fermentation, behaviour and welfare. In experiment 1, eight tethered rumen fistulated Holstein heifers (143 ± 8.7 kg, initial BW) were used to study the effects of treatment on intake, ruminal changes and behaviour. In experiment 2, 48 Friesian female calves (initial BW = 84.5 ± 1.37 kg) were used to study the effects of treatment on performance, behaviour and welfare in feedlot conditions. In both experiments, treatments consisted of concentrate with barley straw (BS) or without barley straw (NBS) and feed was offered at 0830 h on an ad libitum basis. Experiment 1 was performed in four 42-day periods, and sampling was carried out in the last week of each period. Ruminal samples were collected over 3 days at 0, 4, 8 and 12 h post-feeding to measure pH, and volatile fatty acids (VFA) and NH(3)-N concentrations. Maintenance behaviour was video-recorded for 24 h over three consecutive days of each experimental period and feed intake pattern was studied by means of feed bunks mounted on digital platform scales. There were no statistical differences in average daily gain (ADG), concentrate dry matter (DM) intake or CP intake. In contrast, NDF intake and ADF intake were greater in heifers fed BS than NBS. Average ruminal pH was lower, whereas total VFA was greater, in heifers fed NBS diet. There were no differences in NH(3)-N, and in D- and L-lactate concentrations. Time spent in ruminating was shorter, and stereotypies were more frequent in heifers fed diet NBS compared with those fed BS. In experiment 2, nine 28-day periods were established, in which DM intake and ADG were measured, blood and faecal samples were taken for haptoglobin and glucocorticoid metabolites determination, respectively, as welfare indicators, and behaviour was monitored by video recording. Concentrate intake was similar in both treatments, but total feed intake was greater in heifers fed BS diet. As there were no differences in ADG between treatments, gain efficiency was lower in those fed BS than those fed NBS. Blood haptoglobin and faecal glucocorticoids metabolites were not different between treatments. In these competitive conditions, rumination was also reduced and stereotypic behaviour increased by straw exclusion. In conclusion, performance was either not affected or improved by straw exclusion, but animal behaviour was affected, suggesting a negative effect on animal welfare.


HIV, Tuberculosis, and Food Insecurity in Africa-A Syndemics-Based Scoping Review.
The double burden of HIV/AIDS and tuberculosis (TB), coupled with endemic and problematic food insecurity in Africa, can interact to negatively impact health outcomes, creating a syndemic. For people living with HIV/AIDS (PWH), food insecurity is a significant risk factor for acquiring TB due to the strong nutritional influences and co-occurring contextual barriers. We aim to synthesize evidence on the syndemic relationship between HIV/AIDS and TB co-infection and food insecurity in Africa. We conducted a scoping review of studies in Africa that included co-infected adults and children, with evidence of food insecurity, characterized by insufficient to lack of access to macronutrients. We sourced information from major public health databases. Qualitative, narrative analysis was used to synthesize the data. Of 1072 articles screened, 18 articles discussed the syndemic effect of HIV/AIDS and TB co-infection and food insecurity. Reporting of food insecurity was inconsistent, however, five studies estimated it using a validated scale. Food insecure co-infected adults had an average BMI of 16.5-18.5 kg/m2. Negative outcomes include death (n = 6 studies), depression (n = 1 study), treatment non-adherence, weight loss, wasting, opportunistic infections, TB-related lung diseases, lethargy. Food insecurity was a precursor to co-infection, especially with the onset/increased incidence of TB in PWH. Economic, social, and facility-level factors influenced the negative impact of food insecurity on the health of co-infected individuals. Nutritional support, economic relief, and psychosocial support minimized the harmful effects of food insecurity in HIV-TB populations. Interventions that tackle one or more components of a syndemic interaction can have beneficial effects on health outcomes and experiences of PWH with TB in Africa.


Effects of cytokines on rat insulinoma INS-1 cells.
To investigate further the role of cytokines in the pathogenesis of type I insulin-dependent diabetes mellitus, the effects of interleukin-1 beta (IL-1), tumour necrosis factor-alpha (TNF) and gamma-interferon (IFN) were tested on rat insulinoma INS-1 cells. Whereas TNF and IFN had, respectively, a minor or no effect on insulin production, IL-1 caused a time- and dose-dependent decrease in insulin release and lowered the insulin content as well as the preproinsulin mRNA content of INS-1 cells. Both IL-1 and TNF exerted a cytostatic effect, estimated by a decrease in [3H]thymidine incorporation, while only IL-1 decreased cell viability as measured by the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test. The glutathione content of INS-1 cells was shown to be modulated by the presence of 2-mercaptoethanol in the culture medium, but was not affected by IL-1 or TNF. In conclusion, INS-1 cell culture is considered to be a useful model for studying the effect of cytokines on insulin-producing cells. The differentiated features of these cells will permit several questions to be addressed regarding the mechanism of action of IL-1 and eventually other cytokines, both at the level of gene expression and of intracellular signalling.


Spread of Antimicrobial-Resistant Salmonella from Poultry to Humans in Thailand.
Food animal production is important for every country. Several antibiotic agents are used in poultry farming to reduce the economic losses arising from mostly untested infectious diseases. This continued study was performed to determine the prevalence of antibiotic-resistant Salmonella in broiler chickens, poultry farmers, and Salmonella bacteremia patients. A total of 121 Salmonella isolates were collected from the Thai provinces of Khon Kaen (65 isolates), Ratchaburi (43 isolates), and Phayao (13 isolates). Salmonella from chicken showed a high rate of resistance to nalidixic acid and tetracycline. Sixty-four percent of Salmonella isolates carried class 1 integrons (intI1 gene-positive). Among the 121 Salmonella isolates, there were 15 serotypes, with S. Enteritidis being the most common. A clonal relationship between the chicken and human isolates was demonstrated by 3 molecular typing methods: enterobacterial repetitive intergenic consensus polymerase chain reaction; pulsed-field gel electrophoresis; and high-throughput multilocus sequence typing. A spread of the sequence type 11 clone was found between chickens and humans. This study revealed a large-scale Salmonella outbreak in Thailand, a link between resistant bacteria from poultry farms and vertical transmission through the food chain, and horizontal transmission of resistance genes. These results can be used for future surveillance and monitoring.


Oral Health and Quality of Life: Current Concepts.
The recognition of health related quality of life began ever since WHO expanded the definition of health in 1948. This has resulted in the paradigm shift of health and disease from a medical model to a biopsychosocial model. Oral health too denotes not merely the absence of disease but the general well being so that the person can perform functions like eating, talking and smiling and also can contribute creatively to the society. Health related quality of life is a trade-off between how long and how well people live. To explain the concept various theoretical models have been proposed, of which the conceptual model of Wilson and Cleary 1995 is a comprehensive one. Even after much research and thousands of publications the definition of oral health related quality of life is still vague. But the patient's self perception about his oral health and related life quality are significant in clinical dental practice, dental education and research. It is widely shown that oral conditions can have varied impacts on daily living. To assess this, many measures or scales are available. They differ depending on the response format, number of items, context of use and the population in which it is applied. Patient reported outcome assessment is a less developed area in clinical dentistry and research and in future it has the potential to become the primary or secondary outcome measure in clinical interventional research.


THE INFLAMMATORY BOWEL DISEASE-FATIGUE PATIENT SELF-ASSESSMENT SCALE: TRANSLATION, CROSS-CULTURAL ADAPTATION AND PSYCHOMETRIC PROPERTIES OF THE BRAZILIAN VERSION (IBD-F BRAZIL).
Fatigue is a common symptom in patients with inflammatory bowel diseases (IBD). A translated and culturally adapted, instrument with robust psychometric for measuring fatigue in Brazilian patients with IBD is needed. To translate and cross-culturally adapt the inflammatory Bowel Disease Fatigue Scale (IBD-F) into Brazilian-Portuguese and to test its measurement properties in Brazilian patients with IBD. Data from 123 patients with IBD were collected. In addition to IBD-F, the Functional Assessment of Chronic Illness Therapy-Fatigue (FACIT-F) was used. The measurement properties tested were: internal consistency, reproducibility (reliability and agreement), construct validity, internal and external responsiveness, and ceiling and floor effects. The Brazilian-Portuguese version of the IBD-F showed excellent internal consistency (Cronbach's alpha of 0.95), excellent reproducibility (ICC=0.97) and a minimal detectable change of 6.0 points. The construct validity was demonstrated with a good correlation between the IBD-F and FACIT-F (r=- 0.46). Effect sizes used for measuring internal responsiveness were moderate among those with Crohn's (0.66) disease and low in patients with ulcerative colitis (0.24). The Brazilian-Portuguese version of the IBD-F presented with high external responsiveness for Crohn's disease (0.84) and with low external responsiveness for ulcerative colitis (0.33). The area under the curve considered for responsiveness was 0.84. Twenty-five percent of floor effects and no ceiling effect were recorded. The Brazilian-Portuguese version of IBD-F has adequate measurement properties and its use can be recommended in clinical practice and research.


Solvent-polarity dependence of ultrafast excited-state dynamics of trans-4-nitrostilbene.
Ultrafast excited-state dynamics of the simplest nitrostilbenes, namely trans-4-nitrostilbene (t-NSB), was studied in solvents of various polarities with ultrafast broadband time-resolved fluorescence and transient absorption spectroscopies, and by quantum-chemical computations. The results revealed that the initially excited S1(ππ*) state deactivation dynamics is strongly influenced by the solvent polarity. Specifically, the t-NSB S1-state lifetime decreases by three orders of magnitude from ∼60 ps in high-polarity solvents to ∼60 fs in nonpolar solvents. The strong solvent-polarity dependence arises from the differences in dipole moments among the S1 and relevant states, including the major intersystem crossing (ISC) receiver triplet states, and therefore, the solvent polarity can modulate their relative energies and ISC rates. In nonpolar solvents, the sub-100 fs lifetime is due to a combination of efficient ISC and internal conversion. In medium-polarity solvents, the S1-state population decays via a competing ISC relaxation mechanism in a biphasic manner, and the ISC rates are found to obey the inverse energy gap law of the strong coupling case. In high-polarity solvents, the S1 state is stabilized to a much lower energy such that ISC becomes energetically infeasible, and the S1 state decays via barrier crossing along the torsion angle of the central ethylenic bond to the nonfluorescent perpendicular configuration. Regardless of the initial S1-state deactivation pathways in various solvents, the excited-state population is ultimately trapped in the metastable T1-state perpendicular configuration, at which a slower ISC occurs to bring the system to the ground state and bifurcate into either trans or cis form of NSB.


Genetic Determinants of Intrinsic Antibiotic Tolerance in Mycobacterium avium.
The Mycobacterium avium complex (MAC) is one of the most prevalent causes of nontuberculous mycobacteria pulmonary infection in the United States, and yet it remains understudied. Current MAC treatment requires more than a year of intermittent to daily combination antibiotic therapy, depending on disease severity. In order to shorten and simplify curative regimens, it is important to identify the innate bacterial factors contributing to reduced antibiotic susceptibility, namely, antibiotic tolerance genes. In this study, we performed a genome-wide transposon screen to elucidate M. avium genes that play a role in the bacterium's tolerance to first- and second-line antibiotics. We identified a total of 193 unique M. avium mutants with significantly altered susceptibility to at least one of the four clinically used antibiotics we tested, including two mutants (in DFS55_00905 and DFS55_12730) with panhypersusceptibility. The products of the antibiotic tolerance genes we have identified may represent novel targets for future drug development studies aimed at shortening the duration of therapy for MAC infections. IMPORTANCE The prolonged treatment required to eradicate Mycobacterium avium complex (MAC) infection is likely due to the presence of subpopulations of antibiotic-tolerant bacteria with reduced susceptibility to currently available drugs. However, little is known about the genes and pathways responsible for antibiotic tolerance in MAC. In this study, we performed a forward genetic screen to identify M. avium antibiotic tolerance genes, whose products may represent attractive targets for the development of novel adjunctive drugs capable of shortening the curative treatment for MAC infections.


Enrichment programs to create a pipeline to biomedical science careers.
The Student Educational Enrichment Programs at the Medical College of Georgia in the School of Medicine were created to increase underrepresented minorities in the pipeline to biomedical science careers. Eight-week summer programs are conducted for high school, research apprentice, and intermediate and advanced college students. There is a prematriculation program for accepted medical, dental, and graduate students. Between 1979 and 1990, 245 high school students attended 12 summer programs. Of these, 240 (98%) entered college 1 year later. In 1986, after eight programs, 162 (68%) high school participants graduated from college with a baccalaureate degree, and 127 responded to a follow-up survey. Sixty-two (49%) of the college graduates attended health science schools, and 23 (18%) of these matriculated to medical school. Of college students, 504 participated in 13 summer programs. Four hundred (79%) of these students responded to a questionnaire, which indicated that 348 (87%) of the 400 entered health science occupations and/or professional schools; 179 (45%) of these students matriculated to medical school. Minority students participating in enrichment programs have greater success in gaining acceptance to college and professional school. These data suggest that early enrichment initiatives increase the number of underrepresented minorities in the biomedical science pipeline.


Surgery for macular holes associated with unusual concomitant pathologies.
Macular hole (MH) is uncommonly seen in association with retinitis pigmentosa (RP), optic disc pit or choroidal coloboma. The visual disability in eyes with RP, optic disc pit or choroidal coloboma can range from mild to severe. However, the macula may not be involved in the primary pathology and hence, the central vision may be preserved in these eyes. If a MH develops in such eyes the central vision is lost, making the patient severely vision disabled. A perceived poor prognosis in such patients may lead to no intervention. However, an attempt should be made to repair and restore the macula in these situations in order to conserve the vision and limit disability. These eyes can however pose unique surgical challenges. We report our experiences of operating on cases of MH associated with unusual concomitant pathologies such as RP, choroidal coloboma, and optic pit. The outcomes and the surgical challenges differed greatly from the conventional idiopathic MH surgery.


Adaptability and Stability of Faba Bean (Vicia faba L.) Accessions under Diverse Environments and Herbicide Treatments.
The adaptability and stability of 37 faba bean (Vicia faba L.) accessions with different levels of tolerance to metribuzin or imazethapyr was assessed across 12 season-location-herbicide experiments. Significant Genotype x environment (GE) interaction was found for the days to flowering (DFLR), plant height (PLHT) and grain yield (GY). Performance and stability of the accessions regarding PLHT and GY were assessed using four different stability parameters: cultivar superiority, static stability, Wricke's eco-valence and Finlay and Wilkinson's regression model. The stability parameters ranked these genotypes differently suggesting that PLHT and GY stability should be assessed not only on a single or a few stability parameters but on a combination of them. GGE biplot analysis indicated that the environments representing metribuzin treatment at Marchouch 2014-2015 and the non-treated treatment at Terbol 2018-2019 are the ideal environments for evaluating faba bean genotypes. GGE biplots showed herbicide tolerant accession IG12983 with simultaneous average PLHT, GY and stability across the environments. The performance of other tolerant accessions, namely IG13945, IG13906, IG106453, FB2648, and FB1216 was less stable but superior under specific mega environments. Therefore, utilizing these accessions in faba bean breeding programs would help broaden the adaptability to diverse locations-season-herbicide treatments.


Identification and characterization of the immunodominant rat HER-2/neu MHC class I epitope presented by spontaneous mammary tumors from HER-2/neu-transgenic mice.
The HER-2/neu (neu-N)-transgenic mice are a clinically relevant model of breast cancer. They are derived from the parental FVB/N mouse strain and are transgenic for the rat form of the proto-oncogene HER-2/neu (neu). In this study, we report the identification of a MHC class I peptide in the neu protein that is recognized by CD8(+) T cells derived from vaccinated FVB/N mice. This 10-mer was recognized by all tumor-specific FVB/N T cells generated regardless of the TCR Vbeta region expressed by the T cell or the method of vaccination used, establishing it as the immunodominant MHC class I epitope in neu. T cells specific for this epitope were able to cure FVB/N mice of transplanted neu-expressing tumor cells, demonstrating that this is a naturally processed peptide. Altered peptide analogs of the epitope were analyzed for immunogenicity. Vaccination with dendritic cells pulsed with a heteroclitic peptide provided FVB/N and neu-N mice with increased protection against tumor challenge as compared with mice immunized with dendritic cells loaded with either wild-type or irrelevant peptide. Discovery of this epitope allows for better characterization of the CD8(+) T cell responses in the neu-N mouse model in which neu-specific tolerance must be overcome to produce effective antitumor immunity.


Thyroid-stimulating hormone, a novel, locally produced modulator of human epidermal functions, is regulated by thyrotropin-releasing hormone and thyroid hormones.
Several elements of the hypothalamic-pituitary-thyroid axis (HPT) reportedly are transcribed by human skin cell populations, and human hair follicles express functional receptors for TSH. Therefore, we asked whether the epidermis of normal human skin is yet another extrathyroidal target of TSH and whether epidermis even produces TSH. If so, we wanted to clarify whether intraepidermal TSH expression is regulated by TRH and/or thyroid hormones and whether TSH alters selected functions of normal human epidermis in situ. TSH and TSH receptor (TSH-R) expression were analyzed in the epidermis of normal human scalp skin by immunohistochemistry and PCR. In addition, full-thickness scalp skin was organ cultured and treated with TSH, TRH, or thyroid hormones, and the effect of TSH treatment on the expression of selected genes was measured by quantitative PCR and/or quantitative immunohistochemistry. Here we show that normal human epidermis expresses TSH at the mRNA and protein levels in situ and transcribes TSH-R. It also contains thyrostimulin transcripts. Intraepidermal TSH immunoreactivity is up-regulated by TRH and down-regulated by thyroid hormones. Although TSH-R immunoreactivity in situ could not be documented within the epidermis, but in the immediately adjacent dermis, TSH treatment of organ-cultured human skin strongly up-regulated epidermal expression of involucrin, loricrin, and keratins 5 and 14. Thus, normal human epidermis in situ is both an extrapituitary source and (possibly an indirect) target of TSH signaling, which regulates defined epidermal parameters. Intraepidermal TSH expression appears to be regulated by the classical endocrine controls that determine the systemic HPT axis.


Targeting Enclysis in Liver Autoimmunity, Transplantation, Viral Infection and Cancer.
Persistent liver inflammation can lead to cirrhosis, which associates with significant morbidity and mortality worldwide. There are no curative treatments beyond transplantation, followed by long-term immunosuppression. The global burden of end stage liver disease has been increasing and there is a shortage of donor organs, therefore new therapies are desperately needed. Harnessing the power of the immune system has shown promise in certain autoimmunity and cancer settings. In the context of the liver, regulatory T cell (Treg) therapies are in development. The hypothesis is that these specialized lymphocytes that dampen inflammation may reduce liver injury in patients with chronic, progressive diseases, and promote transplant tolerance. Various strategies including intrinsic and extracorporeal expansion of Treg cells, aim to increase their abundance to suppress immune responses. We recently discovered that hepatocytes engulf and delete Treg cells by enclysis. Herein, we propose that inhibition of enclysis may potentiate existing regulatory T cell therapeutic approaches in patients with autoimmune liver diseases and in patients receiving a transplant. Moreover, in settings where the abundance of Treg cells could hinder beneficial immunity, such us in chronic viral infection or liver cancer, enhancement of enclysis could result in transient, localized reduction of Treg cell numbers and tip the balance towards antiviral and anti-tumor immunity. We describe enclysis as is a natural process of liver immune regulation that lends itself to therapeutic targeting, particularly in combination with current Treg cell approaches.


Diazepam attenuation of restraint stress-induced corticosterone levels is enhanced by prior exposure to repeated restraint.
Prior research has demonstrated that diazepam decreases hypothalamic-pituitary-adrenal cortex (HPA) axis activity in stressful contexts but, paradoxically, acts as a stimulator of basal axis activity. Also, several investigators have reported that low doses of diazepam are not effective in reducing stress-induced corticosterone (CORT) levels, yet similar doses typically produce anxiolytic effects on behavioral measures of fear and anxiety. We have examined the effects of diazepam on plasma CORT levels in male Sprague-Dawley rats utilizing a repeated restraint paradigm. Consistent with most literature, diazepam administered IP (1.5, 3.0, or 6.0 mg/kg) 1 h prior to restraint increased non-stress, baseline plasma CORT levels in a dose-dependent fashion. During the first exposure to the 1 h restraint-stress procedure, CORT levels of diazepam-injected rats did not differ from the stress levels of controls except at the 60-min stress time point in those subjects receiving 6.0 mg/kg. However, diazepam at all three doses was able to attenuate the stress-induced increase in CORT following 5 days of diazepam+restraint treatment. Using the 3.0 mg/kg dose as a probe, it was found that this effect was not dependent on the repeated administration of diazepam, but rather on repeated exposure to restraint. These results suggest that repeated restraint produces a change in neural sensitivity to benzodiazepines.


Detection of ECG waveforms by neural networks.
In this study, ECG waveform detection was performed by using artificial neural networks (ANNs). Initially, the R peak of the QRS complex is detected, and then feature vectors are formed by using the amplitudes of the significant frequency components of the DFT spectrum. Grow and Learn (GAL) and Kohonen networks are comparatively investigated to detect four different ECG waveforms. The comparative performance results of GAL and Kohonen networks are reported.


Structural diversity of Burkholderia pseudomallei lipopolysaccharides affects innate immune signaling.
Burkholderia pseudomallei (Bp) causes the disease melioidosis. The main cause of mortality in this disease is septic shock triggered by the host responding to lipopolysaccharide (LPS) components of the Gram-negative outer membrane. Bp LPS is thought to be a weak inducer of the host immune system. LPS from several strains of Bp were purified and their ability to induce the inflammatory mediators TNF-α and iNOS in murine macrophages at low concentrations was investigated. Innate and adaptive immunity qPCR arrays were used to profile expression patterns of 84 gene targets in response to the different LPS types. Additional qPCR validation confirmed large differences in macrophage response. LPS from a high-virulence serotype B strain 576a and a virulent rough central nervous system tropic strain MSHR435 greatly induced the innate immune response indicating that the immunopathogenesis of these strains is different than in infections with strains similar to the prototype strain 1026b. The accumulation of autophagic vesicles was also increased in macrophages challenged with highly immunogenic Bp LPS. Gene induction and concomitant cytokine secretion profiles of human PBMCs in response to the various LPS were also investigated. MALDI-TOF/TOF was used to probe the lipid A portions of the LPS, indicating substantial structural differences that likely play a role in host response to LPS. These findings add to the evolving knowledge of host-response to bacterial LPS, which can be used to better understand septic shock in melioidosis patients and in the rational design of vaccines.


Hypothalamic control systems show differential gene expression during spontaneous daily torpor and fasting-induced torpor in the Djungarian hamster (Phodopus sungorus).
Djungarian hamsters are able to use spontaneous daily torpor (SDT) during the winter season as well as fasting-induced torpor (FIT) at any time of the year to cope with energetically challenging environmental conditions. Torpor is a state of severely reduced metabolism with a pronounced decrease in body temperature, which enables animals to decrease their individual energy requirements. Despite sharing common characteristics, such as reduced body mass before first torpor expression and depressed metabolism and body temperature during the torpid state, FIT and SDT differ in several physiological properties including torpor bout duration, minimal body temperature, fuel utilization and circadian organization. It remains unclear, whether SDT and FIT reflect the same phenomenon or two different physiological states. The hypothalamus has been suggested to play a key role in regulating energy balance and torpor. To uncover differences in molecular control mechanisms of torpor expression, we set out to investigate hypothalamic gene expression profiles of genes related to orexigenic (Agrp/Npy), circadian clock (Bmal1/Per1) and thyroid hormone (Dio2/Mct8) systems of animals undergoing SDT and FIT during different torpor stages. Orexigenic genes were mainly regulated during FIT and remained largely unaffected by SDT. Expression patterns of clock genes showed disturbed circadian clock rhythmicity in animals undergoing FIT, but not in animals undergoing SDT. During both, SDT and FIT, decreased Dio2 expression was detected, indicating reduced hypothalamic T3 availability in both types of torpor. Taken together, our results provide evidence that SDT and FIT also differ in certain central control mechanisms and support the observation that animals undergoing SDT are in energetical balance, whereas animals undergoing FIT display a negative energy balance. This should be carefully taken into account when interpreting data in torpor research, especially from animal models of fasting-induced hypometabolism such as mice.


A novel KCNA1 mutation associated with global delay and persistent cerebellar dysfunction.
Episodic Ataxia Type 1 is an autosomal dominant disorder characterized by episodes of ataxia and myokymia. It is associated with mutations in the KCNA1 voltage-gated potassium channel gene. In the present study, we describe a family with novel clinical features including persistent cerebellar dysfunction, cerebellar atrophy, and cognitive delay. All affected family members have myokymia and epilepsy, but only one individual has episodes of vertigo. Additional features include postural abnormalities, episodic stiffness and weakness. A novel KCNA1 mutation (c.1222G>T) which replaces a highly conserved valine with leucine at position 408 (p.Val408Leu) was identified in affected family members, and was found to augment the ability of the channel to inactivate. Together, our data suggests that KCNA1 mutations are associated with a broader clinical phenotype, which may include persistent cerebellar dysfunction and cognitive delay.


Assessment of ultrasonographic features of polycystic ovaries is associated with modest levels of inter-observer agreement.
There is growing acceptance that polycystic ovaries are an important marker of polycystic ovary syndrome (PCOS) despite significant variability when making the ultrasound diagnosis. To better understand the nature of this variability, we proposed to evaluate the level of inter-observer agreement when identifying and quantifying individual ultrasonographic features of polycystic ovaries. Digital recordings of transvaginal ultrasound scans performed in thirty women with PCOS were assessed by four observers with training in Radiology or Reproductive Endocrinology. Observers evaluated the scans for: 1) number of follicles >/= 2 mm per ovary, 2) largest follicle diameter, 3) ovarian volume, 4) follicle distribution pattern and 5) presence of a corpus luteum (CL). Lin's concordance correlation coefficients and kappa statistics for multiple raters were used to assess inter-observer agreement. Agreement between observers ranged from 0.08 to 0.63 for follicle counts, 0.27 to 0.88 for largest follicle diameter, 0.63 to 0.86 for ovarian volume, 0.51 to 0.76 for follicle distribution pattern and 0.76 to 0.90 for presence of a CL. Overall, reproductive endocrinologists demonstrated better agreement when evaluating ultrasonographic features of polycystic ovaries compared to radiologists (0.71 versus 0.53; p = 0.04). Inter-observer agreement for assessing ultrasonographic features of polycystic ovaries was moderate to poor. These findings support the need for standardized training modules to characterize polycystic ovarian morphology on ultrasonography.


Reporting Maltreatment to Child Protective Services in the Context of Intimate Partner Violence Research.
Given the overlap between intimate partner violence (IPV) and child maltreatment, IPV-exposed child participants in research might disclose instances of child maltreatment. Such disclosures might require researchers to report the maltreatment to child protective services (CPS). However, the literature provides minimal guidance on how to navigate the complex challenges and ethical dilemmas around reporting in the context of research. To help address this gap and stimulate discussion regarding protocols and policies for reporting child maltreatment, this article presents a CPS reporting protocol developed as part of a community-engaged research project evaluating a parenting intervention for system-involved mothers experiencing IPV.


[What does prevention and rehabilitation research contribute to health policy?].
Rational health policy calls for a close collaboration between health-related political organizations and scientific communities, in particular public health sciences and health economics. Based on selected examples, this contribution highlights the benefits of primary and secondary prevention measures with re-spect to two widely distributed chronic diseases, coronary heart disease and depressive disorders. These examples refer to (1) lifestyle changes in secondary prevention of coronary heart disease, (2) theory-based measures of worksite health promotion, and (3)early detection and optimised treatment of depressive disorders. The relevance of interdisciplinary research on prevention and rehabilitation is emphasized by increasing pressures that arise from the current health care system.


Follicular somatic cell factors and follicle development.
Considerable attention is currently paid to oocyte-derived secreted factors that act upon cumulus and granulosa cells. Also important for follicle development are somatic cell-derived secreted factors. This is illustrated by the ability of granulosa cell-derived Kit ligand (KITL) to promote primordial follicle activation, and the loss of follicle development that accompanies KITL gene disruption. This review summarises our current understanding of somatic cell factors during both preantral and antral follicle growth, involving not only signalling from granulosa cells to the oocyte, but also signalling between granulosa and theca cells. Principal granulosa cell-derived factors include activin, anti-Müllerian hormone (AMH), bone morphogenetic proteins (BMPs) and fibroblast growth factors (FGFs). Theca cells also secrete BMPs and FGFs. The interplay between these factors is equally important for follicle growth as the activity of oocyte-derived factors.


Hierarchical assembly of the budding yeast kinetochore from multiple subcomplexes.
Kinetochores are multiprotein complexes that assemble on centromeric DNA and attach chromosomes to spindle microtubules. Over the past six years, the number of proteins known to localize to the Saccharomyces cerevisiae kinetochore has increased from around 10 to over 60. However, relatively little is known about the protein-protein interactions that mediate kinetochore assembly or about the overall structure of microtubule-attachment sites. Here we used biophysical techniques, affinity purification, mass spectrometry, and in vivo assays to examine the state of association of 31 centromere-binding proteins, including six proteins newly identified as kinetochore subunits. We found that yeast kinetochores resemble transcriptional enhancers in being composed of at least 17 discrete subcomplexes that assemble on DNA to form a very large structure with a mass in excess of 5 MD. Critical to kinetochore assembly are proteins that bridge subunits in direct contact with DNA and subunits bound to microtubules. We show that two newly identified kinetochore complexes, COMA (Ctf19p-Okp1p-Mcm21p-Ame1p) and MIND (Mtw1p including Nnf1p-Nsl1p-Dsn1p) function as bridges. COMA, MIND, and the previously described Ndc80 complex constitute three independent and essential platforms onto which outer kinetochore proteins assemble. In addition, we propose that the three complexes have different functions with respect to force generation and MT attachment.


Lipid-binding properties of human ApoD and Lazarillo-related lipocalins: functional implications for cell differentiation.
Lipocalins are a family of proteins characterized by a conserved eight-stranded β-barrel structure with a ligand-binding pocket. They perform a wide range of biological functions and this functional multiplicity must relate to the lipid partner involved. Apolipoprotein D (ApoD) and its insect homologues, Lazarillo (Laz) and neural Lazarillo (NLaz), share common ancestral functions like longevity, stress resistance and lipid metabolism regulation, coexisting with very specialized functions, like courtship behavior. Using tryptophan fluorescence titration, we screened the binding of 15 potential lipid partners for NLaz, ApoD and Laz and uncovered several novel ligands with apparent dissociation constants in the low micromolar range. Retinoic acid (RA), retinol, fatty acids and sphingomyelin are shared ligands. Sterols, however, showed a species-specific binding pattern: cholesterol did not show strong binding to human ApoD, whereas NLaz and Laz did bind ergosterol. Among the lipocalin-specific ligands, we found that ApoD selectively binds the endocannabinoid anandamide but not 2-acylglycerol, and that NLaz binds the pheromone 7-tricosene, but not 7,11-heptacosadiene or 11-cis-vaccenyl acetate. To test the functional relevance of lipocalin ligand binding at the cellular level, we analyzed the effect of ApoD, Laz and NLaz preloaded with RA on neuronal differentiation. Our results show that ApoD is necessary and sufficient to allow for RA differentiating activity. Both human ApoD and Drosophila NLaz successfully deliver RA to immature neurons, driving neurite outgrowth. We conclude that ApoD, NLaz and Laz bind selectively to a different but overlapping set of lipid ligands. This multispecificity can explain their varied physiological functions.


Straight to the point: targeted mRNA-delivery to immune cells for improved vaccine design.
With the deepening of our understanding of adaptive immunity at the cellular and molecular level, targeting antigens directly to immune cells has proven to be a successful strategy to develop innovative and potent vaccines. Indeed, it offers the potential to increase vaccine potency and/or modulate immune response quality while reducing off-target effects. With mRNA-vaccines establishing themselves as a versatile technology for future applications, in the last years several approaches have been explored to target nanoparticles-enabled mRNA-delivery systems to immune cells, with a focus on dendritic cells. Dendritic cells (DCs) are the most potent antigen presenting cells and key mediators of B- and T-cell immunity, and therefore considered as an ideal target for cell-specific antigen delivery. Indeed, improved potency of DC-targeted vaccines has been proved in vitro and in vivo. This review discusses the potential specific targets for immune system-directed mRNA delivery, as well as the different targeting ligand classes and delivery systems used for this purpose.


Accidental mushroom poisoning mimicking stroke. A case report and literature review.
We describe here a paradigmatic case of mushroom poisoning mimicking a stroke. A 64-year old male was referred to the emergency department (ED) for a car accident. He was found diaphoretic, hypotensive, bradycardic, and slightly confused at presentation. No signs of trauma were observed on physical examination. The patient had weakness of the right limbs and bilateral severe myosis. The lab tests were normal, except for leukocytosis, mild hyperglycemia, mild hyperazotemia and moderate hypokalemia. The clinical picture, with the exception of miosis, was thereby suggestive for a stroke, which was also considered the cause of the car accident. The patients' wife, who was brought later to the ED, reported that the husband had suffered a stroke 4 years earlier, with residual right hemiparesis. She also referred that the patient showed signs of diaphoresis and confusion, and the car was intentionally driven into the ditch. Among other details, she referred that the husband ate mushrooms that she had personally collected 2 hours before taking the drive. Two mg of atropine, intravenous rehydration and potassium were hence administered in the suspicion of a cholinergic toxydrome, and complete clinical remission was rapidly obtained. Among the mixture of mushrooms eaten by the patient, a mycologist identified Armillaria Mellea (an edible mushroom) and notably Inocybe Fastigiata, a toxic muscarine-containing mushroom, easily confounded with Armillaria. After observation and oral rehydration, the patient was discharged.


[The effect of exogenous DNA on the bodies and cells of eukaryotes].
A review of literature and the authors own data on specific and non-specific effect of DNA on the organism and eucaryote cells are presented. A conclusion is made that transduction and transformation in eucaryotes might be possible. It is confirmed by a great number of experimental data. The problem on mutagenic effect of DNA still remains disputable. The problem of DNA restriction in eucaryotes is absolutely unstudied and requires experimental development. A great theoretical and practical significance of the studies in the effect of exogenic DNA in higher organisms is emphasized.


Immunoinhibitory checkpoint deficiency in medium and large vessel vasculitis.
Giant cell arteritis (GCA) causes autoimmune inflammation of the aorta and its large branches, resulting in aortic arch syndrome, blindness, and stroke. CD4+ T cells and macrophages form organized granulomatous lesions in the walls of affected arteries, destroy the tunica media, and induce ischemic organ damage through rapid intimal hyperplasia and luminal occlusion. Pathogenic mechanisms remain insufficiently understood; specifically, it is unknown whether the unopposed activation of the immune system is because of deficiency of immunoinhibitory checkpoints. Transcriptome analysis of GCA-affected temporal arteries revealed low expression of the coinhibitory ligand programmed death ligand-1 (PD-L1) concurrent with enrichment of the programmed death-1 (PD-1) receptor. Tissue-residing and ex vivo-generated dendritic cells (DC) from GCA patients were PD-L1lo, whereas the majority of vasculitic T cells expressed PD-1, suggesting inefficiency of the immunoprotective PD-1/PD-L1 immune checkpoint. DC-PD-L1 expression correlated inversely with clinical disease activity. In human artery-SCID chimeras, PD-1 blockade exacerbated vascular inflammation, enriched for PD-1+ effector T cells, and amplified tissue production of multiple T-cell effector cytokines, including IFN-γ, IL-17, and IL-21. Arteries infiltrated by PD-1+ effector T cells developed microvascular neoangiogenesis as well as hyperplasia of the intimal layer, implicating T cells in the maladaptive behavior of vessel wall endogenous cells. Thus, in GCA, a breakdown of the tissue-protective PD1/PD-L1 checkpoint unleashes vasculitic immunity and regulates the pathogenic remodeling of the inflamed arterial wall.


Symptoms and clinical and radiological signs predicting the presence of pathogenic bacteria in acute rhinosinusitis.
A minority of patients with upper respiratory tract infections (URTI) have a bacterial infection and may benefit from antibiotherapy. In previous investigations we showed that in patients suffering from acute rhinosinusitis associated with the presence of Streptococcus pneumoniae, Haemophilus influenzae or Moraxella catarrhalis in their nasopharygeal secretions, resolution of symptoms was significantly improved by antibiotic treatment. The present analysis was performed to determine whether specific clinical symptoms or signs observed during careful endoscopic examination of the nasal cavities could help the clinician to identify a subset of patients with moderate forms of acute rhinosinusitis infected with pathogenic bacteria. Detailed clinical histories were obtained and medical examinations performed in 265 patients (138 females, 127 males; mean age 35 years) presenting with a < 4-week history of URTI symptoms but who did not require immediate antibiotic therapy for severe rhinosinusitis. The presence of three pathogenic bacteria (S. pneumoniae, H. influenzae and M. catarrhalis) was determined in all patients by culture of nasopharyngeal secretions. Azithromycin (500 mg/day for 3 days; n = 133) or placebo (n = 132) were randomly given to all patients in a double-blind manner. Pathogenic bacteria were found in 77 patients (29%). The clinical signs and symptoms significantly associated in a multivariate model with the presence of bacteria included colored nasal discharge (p < 0.003), facial pain (p < 0.032) and radiologically determined maxillary sinusitis (complete opacity, air-fluid level or mucosal thickening > 10 mm) (p < 0.001). This best predictive model had a sensitivity of 69% and a specificity of 64% and therefore could not be used either as a screening tool or as a diagnostic criterion for bacterial rhinosinusitis. In the group of patients with positive bacterial cultures, resolution of symptoms at Day 7 was observed in 73% of patients treated with azithromycin and in 47% of patients in the placebo group (p < 0.007). We conclude that signs and symptoms of acute rhinosinusitis in patients with mild-to-moderate clinical presentations are poor predictors of the presence of bacteria.


RABIES IN RODENTS AND LAGOMORPHS IN THE USA, 2011-20.
Rabies is an acute progressive encephalitis caused by infection with rabies viruses, with reservoirs among bats and mesocarnivores, but all mammals are susceptible. Despite its distribution and abundance, cases of rabies are much less common in rodents and lagomorphs. Familiarity with current rabies prevalence data is important for informed decisions on human postexposure prophylaxis after rodent and lagomorph bites. This study is an update of rabies cases reported in rodents and lagomorphs in the US from 2011 to 2020. Rabies reports were collected passively from laboratory testing agencies in the US and Puerto Rico from 2011 to 2020. Descriptive analysis was conducted to determine the percent positivity of rabies cases by species. A total of 401 cases of rabies in rodents and lagomorphs were reported from 2011 to 2020. Most reported cases were in groundhogs (Marmota monax), representing >90% of cases, and the trend closely aligned with rabies in raccoons (Procyon lotor). In any given year, the percent positivity of rabies in rodents and lagomorphs was <2.5%, and the trend of percent positivity from 2011 to 2020 was stable. Groundhog and North American beaver (Castor canadensis) percent positivity was significantly higher than the rest of the rodents and lagomorphs. Most rabies cases occurred during the months of May-September. Documented cases of rabies in rodents and lagomorphs are generally rare, but with variation between species. Groundhogs and North American beavers had rabies percent positivity similar to high-risk species, such as bats and raccoons, and constituted 97% of all rodent and lagomorph positive cases. Since 1993, the trend in rabies cases in groundhogs has significantly declined. These results can be used to help inform public health officials on rodent and lagomorph prevention and control efforts, as well as rabies postexposure prophylaxis.


[Effects of KS-R1 (ampicillin suppository) and ampicillin oral dosing on fecal flora of children].
A newly developed product, KS-R1 (a suppository containing 250 mg potency of ampicillin (ABPC)), was given to 7 children (5 boys and 2 girls) ranging from 4 years and 5 months to 8 years and 10 months in age, 3 times a day (average daily dose, 62.4 mg/kg) for 5 days. As a control, the same amount of ABPC dry syrup was given orally to 7 children (4 boys and 3 girls) ranging from 2 years and 8 months to 7 years and 7 months in age, 3 times a day (average daily dose, 55.3 mg/kg) for 5 days. The effects of these 2 preparations on the bacterial flora of the feces were investigated, and concentrations of ABPC in the feces and sensitivities of the isolated strains to ABPC were determined. The results were as follows. As for Gram-negative bacilli in the feces from children given KS-R1, there was no change in Escherichia coli, Klebsiella sp., Citrobacter sp. and Enterobacter sp. which were isolated from many children 3 days after the end of treatment, but they did not show any constant pattern of change in mean number. Other species did not show any pattern of increase in the number of children from whom they were isolated, or in the number of bacteria during the course after the beginning of treatment. The total number of bacteria identified as Enterobacteriaceae was at the level of 10(8) cells/g on any day of examination. Of Gram-negative bacilli other than Enterobacteriaceae, Pseudomonas sp. showed no constant pattern of change in number. On the other hand, of Gram-positive bacteria, there was no constant pattern of change in the number of Staphylococcus aureus, Coagulase-negative Staphylococci began to be isolated from many children 5 days after the beginning of treatment and were isolated from all children 5 days after the end of treatment, but there was no tendency for the mean number to increase. Enterococcus sp. was not isolated from 3 children 3 days after the beginning of treatment, and was decreased in number by one order in 3 out of other 4 children as compared with before-treatment. However, this species was isolated from all children 3 days after the end of treatment, and the mean number of bacteria was similar to that before-treatment although the number of bacteria was different in individual children.(ABSTRACT TRUNCATED AT 400 WORDS)